Chem. J. Chinese Universities ›› 2009, Vol. 30 ›› Issue (7): 1410.

• Articles • Previous Articles     Next Articles

Specific-site PEGylation of Hirudin on Ion-exchange Column and Theoretical Prediction and Analysis of Modified Sites by Molecular Dynamics Simulation

ZHAO Jun, LI Xue-Qin, XIU Zhi-Long*   

  1. Department of Bioscience and Biotechnology, Dalian University of Technology, Dalian 116024, China
  • Received:2008-03-28 Online:2009-07-10 Published:2009-07-10
  • Contact: XIU Zhi-Long. E-mail: zhlxiu@dlut.edu.cn
  • Supported by:

    国家自然科学基金(批准号: 20176005)资助.

Abstract:

Hirudin, a remarkably stable molecule with blood anticoagulant activity, is the most potent thrombin inhibitor in nature. But the short half life in blood plasma limits the application of hirudin in medical care. PEGylation is widely used to prolong the half life of proteins and peptides in plasma. However, the potential PEGylation sites of hirudin are numerous and it is difficult to confirm the actual PEGylation sites. In this paper, based on our experiment results of PEGylated hirudin in solution phase PEGylation and on-column PEGylation, hirudin and hirudin/thrombin complex had been simulated using molecular dynamics to predict the Lys PEGylation sites of hirudin. The results show that during the liquid-phase PEGylation, lysine residues are easy to be PEGylated except for Lys47, while during the solid-phase PEGylation, only Lys35 and Lys27 are easy to be PEGylated. And the PEGylation product can be stable when Lys35 is chosen to be the PEGylation site.

Key words: Hirudin; Molecular dynamics simulation; PEGylation; Solvent accessible surface

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